Human antibody-producing cell

Abstract

A chicken B cell that expresses a variety of human antibodies. A chicken B cell, in which, in an antibody light chain gene locus thereof, all or a part of a DNA sequence derived from a human antibody light chain variable region and a human antibody light chain constant region are inserted, or the antibody light chain gene locus is replaced with all or a part of a DNA sequence derived from human antibody light chain variable and constant regions, and in an antibody heavy chain gene locus thereof, all or a part of a DNA sequence derived from human antibody heavy chain variable and constant regions are inserted, or the antibody heavy chain gene locus is replaced with all or a part of a DNA sequence derived from human antibody heavy chain variable and constant regions, and in an antibody light chain pseudogene locus thereof, two or more DNA sequences derived from human antibody light chain variable regions are inserted, or the antibody light chain pseudogene locus is replaced with two or more DNA sequences derived from human antibody light chain variable regions, and/or in an antibody heavy chain pseudogene locus thereof, two or more DNA sequences derived from human antibody heavy chain variable regions are inserted, or the antibody heavy chain pseudogene locus is replaced with two or more DNA sequences derived from human antibody heavy chain variable regions.

1 . (canceled) 
     
     
         2 . A chicken B cell, in which the chicken antibody light chain gene locus is replaced by all or a part of a DNA sequence derived from a human antibody light chain variable region and by all or a part of a DNA sequence derived from a human antibody light chain constant region,
 which in the chicken antibody heavy chain gene locus all or a part of a DNA sequence derived from a human antibody heavy chain variable region and of a human antibody heavy chain constant region are inserted,   and in which the chicken antibody light chain pseudogene locus is replaced by 15 or more DNA sequences derived from human antibody light chain variable regions,   and in which the chicken antibody heavy chain pseudogene locus is inserted by 15 or more DNA sequences derived from human antibody heavy chain variable regions,   wherein the chicken B cell has the ability to express a human antibody on the cell surface and also to secrete the human antibody into the culture solution, and wherein a complementarity-determining region 3 (CDR3) encoded by the human antibody light chain and heavy chain variable regions comprises more than 5 different amino acids.   
     
     
         3 . The chicken B cell according to  claim 2 , in which the chicken antibody light chain pseudogene locus is replaced by DNA sequence comprising SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, and SEQ ID NO:88,
 and in which the chicken antibody heavy chain pseudogene locus is inserted by DNA sequence comprising SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, SEQ ID NO:106, SEQ ID NO:107, and SEQ ID NO:108.   
     
     
         4 . The chicken B cell according to  claim 2 , in which the chicken antibody light chain pseudogene locus is replaced by DNA sequence comprising SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, and SEQ ID NO:88,
 and in which the chicken antibody heavy chain pseudogene locus is inserted by DNA sequence comprising SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, SEQ ID NO:122, SEQ ID NO:123, and SEQ ID NO:124.   
     
     
         5 . The chicken B cell according to  claim 2 , and in which the chicken antibody light chain pseudogene locus is replaced by DNA sequence comprising SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:139, SEQ ID NO:140, and SEQ ID NO:141,
 and in which the chicken antibody heavy chain pseudogene locus is inserted by DNA sequence comprising SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, SEQ ID NO:122, SEQ ID NO:123, and SEQ ID NO:124.   
     
     
         6 . The chicken B cell according to  claim 2 , in which the chicken antibody light chain pseudogene locus is replaced by DNA sequence comprising SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:139, SEQ ID NO:140, and SEQ ID NO:141,
 and in which the chicken antibody heavy chain pseudogene locus is inserted by DNA sequence comprising SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, SEQ ID NO:122, SEQ ID NO:123, SEQ ID NO:124, SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, SEQ ID NO:154, SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, SEQ ID NO:158, SEQ ID NO:159, SEQ ID NO:160, and SEQ ID NO:161.   
     
     
         7 . The chicken B cell according to  claim 2 , wherein the DNA sequence derived from the human antibody light chain variable region inserted into the antibody light chain gene locus is at a position upstream from the DNA sequence derived from the human antibody light chain constant region, and the DNA sequence derived from the human antibody heavy chain variable region inserted into the antibody heavy chain gene locus is at a position upstream from the DNA sequence derived from the human antibody heavy chain constant region. 
     
     
         8 . The chicken B cell according to  claim 2 , wherein the human antibody heavy chain is a γ chain. 
     
     
         9 . The chicken B cell according to  claim 2 , wherein the human antibody light chain is a λ chain or a κ chain. 
     
     
         10 . The chicken B cell according to  claim 2 , which is a DT40 cell. 
     
     
         11 . The chicken B cell according to  claim 2 , which has been subjected to a treatment of relaxing chromatin. 
     
     
         12 . The chicken B cell according to  claim 11 , wherein the treatment of relaxing chromatin is reduction or deletion of the function of histone deacetylase in the chicken B cell. 
     
     
         13 . The chicken B cell according to  claim 12 , wherein the method for reducing or deleting the function of histone deacetylase is reduction or deletion of the expression of a histone deacetylase gene in the chicken B cell. 
     
     
         14 . The chicken B cell according to  claim 13 , wherein the histone deacetylase is HDAC2. 
     
     
         15 . The chicken B cell according to  claim 12 , wherein the method for reducing or deleting the function of histone deacetylase is a treatment with an HDAC inhibitor. 
     
     
         16 . An antibody-producing cell library consisting of the chicken B cell according to  claim 2 . 
     
     
         17 . A method for producing a human antibody or a humanized antibody from the chicken B cell according to  claim 2 , comprising culturing said chicken B cell under culturing conditions suitable for the cell to produce a human antibody or a humanized antibody and obtaining said antibody. 
     
     
         18 . Antibodies reacting with various antigens, wherein said antibodies have been produced by the cell library according to  claim 16 . 
     
     
         19 . A kit for producing the chicken B cells according to  claim 2 . 
     
     
         20 . The method of  claim 17 , wherein the antibody is comprised in a pharmaceutical product.